Defining a stromal niche for type 2-like lung regulatory T cells

定义 2 型肺调节性 T 细胞的基质生态位

基本信息

批准号：
10536569
负责人：
Anthony Chang
金额：
$ 3.86万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10536569
关键词：
3-Dimensional Ablation Acute Lung Injury Acute Respiratory Distress Syndrome Adipose tissue Biological Response Modifiers Biology CD4 Positive T Lymphocytes Cells Coculture Techniques Data Set Development FOXP3 gene Fibroblasts Flow Cytometry Foundations GATA3 gene Genetic Goals Homeostasis Human Image Immune Immune response In Vitro Inflammation Inflammatory Influenza A virus Injury Integrins Ligands Location Lung Lung diseases Lymphocyte Lymphoid Maintenance Measurement Mediating Microscopy Modeling Morbidity - disease rate Mus Outcome Pathway interactions Play Positioning Attribute Precision therapeutics Production Proliferation Marker Pyroglyphidae Recovery Regulation Regulatory T-Lymphocyte Reporter Role Signal Transduction Stromal Cells Structure of parenchyma of lung Surface System T-bet protein TSLP gene Testing Th2 Cells Therapeutic Uses Thick Tissues Vascular Cell Adhesion Molecule-1 Visceral Work allergic airway disease design differential expression epithelium regeneration experimental study high dimensionality immunoregulation insight intercellular cell adhesion molecule lung injury lung repair mesenchymal stromal cell mortality mouse model novel pulmonary function receptor reparative process response single-cell RNA sequencing transcriptome sequencing

项目摘要

Project Summary Lung diseases such as acute lung injuries (ALI) and acute respiratory distress syndrome (ARDS) are leading causes of morbidity and mortality worldwide. Regulatory T cells (Tregs) are traditionally thought of as critical negative regulators of systemic immune responses; however, their local roles in tissues such as the lung are being increasingly appreciated, where they can promote lung epithelial regeneration in both ALI and ARDS. Subsets of tissue-resident Tregs (tTregs) are found to express transcription factors T-bet, Gata3, RORt, and Bcl6, and have enhanced suppression towards their Th1, Th2, Th3/17, and Tfh immune ‘flavors,’ respectively. Although tTreg function are beginning to be understood, how lung tTregs are regulated, positioned, and maintained within their respective tissue niches remains unknown. Mesenchymal stromal cells (MSCs) are immune regulators and have been highlighted to play a role in tTreg biology. Production of IL-33 by a subset of MSCs promotes the expansion and maintenance of visceral adipose tissue tTregs and human MSCs can induce Tregs from conventional CD4+ T cells in vitro. Our group identified a stromal cell niche within the lung where adventitial fibroblasts (AFs), an MSC subset, regulate type 2 effector lymphocytes (e.g. ILC2s and Th2 cells), in part via the secretion of IL-33 and thymic stromal lymphopoietin (TSLP). Using 3D thick section imaging, I have shown that lung type 2-like tTregs (i.e. Gata3hi ST2+, KLRG1+) also localize to this niche, indicating AFs may regulate lung tTregs and their subsets. When co-cultured with lung AFs, lymphoid Tregs significantly increased proliferation, survival, and expression of type 2-like Treg markers ST2 and KLRG1 in a contact-dependent manner. Additionally, AFs preferentially support ST2hi Tregs, as evidenced by higher proliferation, survivability, and ST2 and KLRG1 expression. Using CellphoneDB V2.0, I identified extracellular matrix (ECM)–integrin ligand–receptor pairings, such as ICAM, VCAM, and CD49d, that may mediate interactions between AFs and Tregs. Upon blocking all three in a co-culture system, I found a significant decrease in Treg proliferation and ST2 and KLRG1 expression. I hypothesize that AFs regulate the maintenance and differentiation of lung Treg subsets, preferentially supporting type 2-like lung Tregs, which play critical roles in post-injury lung repair. This proposal will define the topography of type 2 lung tTregs and their role in naïve and inflammatory settings (Aim 1) and determine the role of adventitial fibroblasts in the regulation of lung Treg subsets (Aim 2). This work utilizes murine models of inflammation, as well as genetic ablations to dissect the local type 2 tTreg response in the lung. High-dimensional flow cytometry, advanced imaging, and lung function measurements will be used to quantify impacts on immune cells and functional lung recovery. Completion of these aims will elucidate the role of AFs in the function and regulation of lung type 2 tTregs, providing novel mechanistic insight into the role MSCs play in regulating immune subsets. Completion of this study provides a foundation for the development of precision therapeutics to selectively regulate lung tissue Tregs subsets to impact the outcome of diverse lung diseases.

项目概要急性肺损伤 (ALI) 和急性呼吸窘迫综合征 (ARDS) 等肺部疾病是主要原因传统上认为调节性 T 细胞 (Treg) 是导致全球发病和死亡的关键因素。然而，它们在肺等组织中的局部作用是它们可以促进 ALI 和 ARDS 中的肺上皮再生，因此受到越来越多的重视。组织驻留性 Tregs (tTreg) 子集被发现表达转录因子 T-bet、Gata3、RORt 和 Bcl6 和分别增强了对其 Th1、Th2、Th3/17 和 Tfh 免疫“风味”的抑制。尽管人们开始了解 tTreg 功能，但肺 tTreg 是如何调节、定位和释放的？间充质基质细胞（MSC）是否维持在各自的组织生态位中仍然未知。免疫调节剂，并已被强调在 tTreg 生物学中发挥作用，由 tTreg 的一个子集产生。 MSC 促进内脏脂肪组织 tTreg 的扩增和维持，人类 MSC 可以诱导我们的研究小组在体外鉴定了来自传统 CD4+ T 细胞的 Treg 细胞。外膜成纤维细胞 (AF) 是 MSC 的一个子集，可调节 2 型效应淋巴细胞（例如 ILC2 和 Th2 细胞）部分通过 IL-33 和胸腺基质淋巴细胞生成素 (TSLP) 的分泌，使用 3D 厚切片成像，我得到了。显示肺 2 型样 tTregs（即 Gata3hi ST2+、KLRG1+）也定位于该生态位，表明 AF 可能调节肺 tTreg 及其亚群当与肺 AF 共培养时，淋巴 Treg 显着增加。接触依赖性细胞中 2 型 Treg 标记物 ST2 和 KLRG1 的增殖、存活和表达此外，AF 优先支持 ST2hi Tregs，这通过较高的增殖能力、生存能力、使用 CellphoneDB V2.0，我鉴定了细胞外基质 (ECM)-整合素。配体-受体配对，例如 ICAM、VCAM 和 CD49d，可能介导 AF 和在共培养系统中阻断所有这三种 Treg 后，我发现 Treg 增殖和 ST2 显着减少。和 KLRG1 表达，AFs 调节肺 Treg 亚群的维持和分化，优先支持 2 型肺 Tregs，其在损伤后肺修复中发挥关键作用。将定义 2 型肺 tTreg 的拓扑结构及其在幼稚和炎症环境中的作用（目标 1）和确定外膜成纤维细胞在肺 Treg 亚群调节中的作用（目标 2）。小鼠炎症模型以及基因消融来剖析肺部局部 2 型 tTreg 反应。高维流式细胞术、先进成像和肺功能测量将用于量化对免疫细胞和功能性肺恢复的影响的完成将阐明 AF 在其中的作用。肺 2 型 tTreg 的功能和调节，为 MSC 的作用提供新的机制见解这项研究的完成为精准化的发展奠定了基础。选择性调节肺组织 Tregs 亚群以影响多种肺部疾病结果的疗法。